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The following results have been obtained in consequence of patho-morphological examination regarding the supplemental groove. 1. Light microscopic observation of cross-sectioned supplemental grooves revealed that most of them were shallow in the form of plate or bowl. Some of the supplemental grooves had contents not described in the past and the structure of the contents was not clear under a light microscope. The contents were found in 22% of the supplemental grooves examined. 2. The contents in supplemental grooves which were confirmed under a light microscope were found to consist of enamel itself when examined by means of an electron microscope. Microhardness measurements of this enamel showed less than one third the values of normal enamel. By means of microradiography, it was established that radiolucency of this enamel was, for the most part, much higher than normal enamel. 3. It was ascertained that enamel with low hardness and high radiolucency constitutes the contents of supplemental grooves. Judging from its tissue properties, the contents were believed to be susceptible to attack by caries. This view was supported by the results of an investigation of caries sites in supplemental grooves.